东海1号大黄鱼SCD-1a和SCD-1b基因克隆及其对低温胁迫的应答  

Gene Cloning of SCD-1a and SCD-1b in Donghai No. 1 Large Yellow Croaker(Larimichthys crocea) and Their Responses to Cold Stress

作　　者：李明云[1] 苗亮[1] 陈莹莹[1] 李翀 张立宁[2] LI Ming-Yun;MIAO Liang;CHEN Ying-Ying;LI Chong;ZHANG Li-Ning(State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products,Ningbo University,Ningbo 315211,China;Zhejiang Mariculture Research Institute,Wenzhou 325005.China)

机构地区：[1]宁波大学农产品质量安全危害因子与风险防控国家重点实验室,宁波315211 [2]浙江省海洋水产养殖研究所,温州325005

出　　处：《农业生物技术学报》2020年第10期1820-1829,共10页Journal of Agricultural Biotechnology

基　　金：浙江省水产育种重大科技专项(2016C02055-7);国家高技术研究发展计划(863计划)(2012AA10A403-4);温州市农业新品种选育协作组项目(2019ZX002)。

摘　　要：大黄鱼(Larimichthys crocea)是暖温性鱼类,冬季低温可引起养殖大黄鱼大量死亡,有必要开展大黄鱼应对低温胁迫的研究。为明确大黄鱼硬脂酰辅酶A去饱和酶1基因(stearoyl coenzyme A desaturase 1, SCD-1)序列特征及其对低温胁迫应激的响应,本研究运用cDNA末端快速扩增(rapid amplification of cDNA ends,RACE)技术从东海1号大黄鱼中克隆获得SCD-1a和SCD-1b的cDNA序列全长,并通过qRT-PCR分析二者在慢性和急性低温胁迫下的表达变化。SCD-1a cDNA (GenBank No. MT325796)全长3 095 bp,开放阅读框1014 bp,编码337个氨基酸;SCD-1b cDNA (GenBank No. MT325797)全长2 636 bp,开放阅读框1 008 bp,编码335个氨基酸。序列比对显示,大黄鱼SCD-1a和SCD-1b基因编码的氨基酸序列相似性为72.2%,都含有3个保守的组氨酸富含区域;二者预测的蛋白质三级结构高度相似,均有4个跨膜螺旋。系统进化分析显示,鱼类SCD聚为2个大簇,大黄鱼SCD-1a和SCD-1b分别位于两簇中,表明二者为不同亚型。正常生长温度(18℃)下,SCD-1a和SCD-1b都在大黄鱼肝脏中高表达,在其他组织中弱表达。慢性低温胁迫(以0.5℃/12 h的速度降温、维持12 h后继续降温的方法由12℃缓慢降至6℃)条件下,鳃、皮肤、肌肉、肠和脑中SCD-1a表达量显著升高,肝中SCD-1a表达量显著降低,皮肤、肌肉、肠、脑和心脏中SCD-1b表达量显著升高;急性低温胁迫(将鱼由12℃池直接捞入8℃池)条件下,所有组织中SCD-1b均出现表达显著升高,SCD-1a在皮肤以外的组织均表达显著升高。本研究结果对于了解大黄鱼应对和适应低温胁迫的机制具有一定的参考意义。Large yellow croaker(Larimichthys crocea) is a kind of warm-temperature fish. Low temperature in winter can cause large death of cultured L. crocea, so it is necessary to carry out researches on how to deal with low temperature stress. In order to study the sequence characteristics of stearoyl coenzyme A desaturase1(SCD-1) gene of Donghai No. 1 large yellow croaker and their responses to cold stress, the full length of SCD-1 a(GenBank No. MT325796) and SCD-1 b(GenBank No. MT325797) cDNA were cloned by rapid amplification of cDNA ends(RACE) technique, and expression level under chronic and acute cold stress were analysed by q RT-PCR. The full length of SCD-1 a cDNA was 3 095 bp which contained a 1 014 bp ORF and encoded 337 amino acid. The full length of SCD-1 b cDNA was 2 636 bp which contained a 1 008 bp ORF and encoded 335 amino acid. SCD-1 a and SCD-1 b both contained 3 conserved histidine rich regions, and the sequence similarity was 72.2%. The predicted tertiary protein structure of SCD-1 a and SCD-1 b were highly similar and both contained 4 transmembrane helical structure. In the phylogenetic tree based on amino acids, the SCD of teleost divided into 2 clusters, the SCD-1 a and SCD-1 b of L. crocea located in 2 clusters separately,indicating that they were different subtypes. Under normal growth temperature(18 ℃), the expression levels of SCD-1 a and SCD-1 b were both high in liver and lower in other tissues. During chronic-cold stress(decreasing slowly from 12 ℃ to 6 ℃ by the method of cooling at the rate of 0.5 ℃/12 h and continuing to cool down after maintaining for 12 h), the expression of SCD-1 a significantly increased in gill, skill, muscle, intestine and brain, and the expression of SCD-1 b significantly increased in skill, muscle, intestine, brain and heart, nevertheless SCD-1 a expression decreased significantly in liver. During acute cold stress(from 12 to 8 ℃ immediately), the expression level of SCD-1 b significantly increased in all tissues, and SCD-1 a expression significantly incr

关 键 词：东海1号大黄鱼 硬脂酰辅酶A去饱和酶1基因(SCD-1) 低温胁迫 组织应答 

分 类 号：S965.3[农业科学—水产养殖] Q785[农业科学—水产科学]